**3.3 Speech synthesis: SSML 1.0 and 1.1**

Another effort was to define how to control a speech synthesis, or TTS engine. This is to help the engine render the textual prompt in the most accurate way. The XML markup language for this purpose is the Speech Synthesis Markup Language Version 1.0, SSML 1.0 [20], which was released in March 2004.

**Figure 4** shows the five major processing steps present in all TTS engines. For each of them, the engine offers a normal behavior, called "non-markup behavior" in the picture. The SSML mark-up instead allows the engine to improve the default rendering by means of elements of the language. Each element is related to one specific processing step, and it is interpreted as a request by the author to perform some specific processing. It is then up to the processor to determine whether and in what way to realize the command.

The SSML example in **Figure 5** shows a prompt for a flight information system structured into a single paragraph (<p > element) and two sentences (<s > elements). Acronyms are substituted (<sub>) into expanded versions, pauses are added (<break>), and a time expression is explicitly labeled (<say-as>) to select the correct way of reading it. Other elements can change additional features, such as prosodic features of speed and rate (<prosody>), and how to change the speaking voice (<voice>).

SSML 1.0 [21] continued to be standardized to promote the use of SSML to more international languages, in particular Asian and Indian languages. Three workshops were held to encourage local companies and universities to propose features to be added to the language:

A new standard SSML 1.1 [24] was released in September 2010. See Appendix F of [24] for details on the changes. Among them, a < token> element was introduced for languages where the whitespace has peculiar behavior, such as in Mandarin, Japanese, Thai, Vietnamese, and Urdu.

**55**

*Speech Standards: Lessons Learnt*

*DOI: http://dx.doi.org/10.5772/intechopen.93134*

**3.4 Pronunciation lexicon: PLS 1.0**

**Figure 5.**

*A simple SSML document.*

will be taken into account.

**3.5 Call control—CCXML 1.0**

or for a different spelling for the same pronunciation.

Both speech grammars and synthesized prompts can require customizing the pronunciation of words in a specific application domain. This is often done by adding a user lexicon. The Pronunciation Lexicon Specification (PLS 1.0 [25]) was created to support the definition of a standard lexicon fully interoperable with SRGS 1.0 and SSML 1.0/1.1. PLS 1.0 became a W3C Recommendation in October 2008. A PLS document is a container of entries, <lexeme> elements, with a textual part described by the <grapheme> element and with textual replacements provided by <alias> elements or phonetic transcriptions by <phoneme> elements. There can be multiple pronunciations to accommodate different ways to speak a word/token,

A simple PLS 1.0 document example for a flight application is shown in **Figure 6**. For "Alitalia" and "Lufthansa," the pronunciations inside the <phoneme> element are given in IPA (International Phonetic Alphabet) [34]—a standard way to express the pronunciations for all spoken human languages. Moreover, the two lexemes have a double pronunciation; the first is the normal English one, while the second is closer to their original language (Italian and German, respectively) as spoken by a native speaker of that language. The prefer attribute indicates which pronunciation has to be selected for TTS rendering. For ASR, all the pronunciations

Another language defined by the W3C VBWG targets programming the call control of a voice browser in an innovative way. An XML markup language was developed to define handlers for telephony events generated by a telephone connection or a VoIP SIP interaction. The Voice Browser Call Control (CCXML 1.0) [26] language was designed to allow a very efficient implementation completely based upon events and handlers to avoid creating any latency that might impact the underlying signaling.

**Figure 4.** *SSML support for stages of speech synthesis.*

*Speech Standards: Lessons Learnt DOI: http://dx.doi.org/10.5772/intechopen.93134*

*Human 4.0 - From Biology to Cybernetic*

what way to realize the command.

voice (<voice>).

added to the language:

• Nov 2005 at Beijing (China)

• May 2006 at Crete (Greece)

Japanese, Thai, Vietnamese, and Urdu.

*SSML support for stages of speech synthesis.*

• Jun 2007 at Hyderabad

**3.3 Speech synthesis: SSML 1.0 and 1.1**

Another effort was to define how to control a speech synthesis, or TTS engine. This is to help the engine render the textual prompt in the most accurate way. The XML markup language for this purpose is the Speech Synthesis Markup Language

**Figure 4** shows the five major processing steps present in all TTS engines. For each of them, the engine offers a normal behavior, called "non-markup behavior" in the picture. The SSML mark-up instead allows the engine to improve the default rendering by means of elements of the language. Each element is related to one specific processing step, and it is interpreted as a request by the author to perform some specific processing. It is then up to the processor to determine whether and in

The SSML example in **Figure 5** shows a prompt for a flight information system structured into a single paragraph (<p > element) and two sentences (<s > elements). Acronyms are substituted (<sub>) into expanded versions, pauses are added (<break>), and a time expression is explicitly labeled (<say-as>) to select the correct way of reading it. Other elements can change additional features, such as prosodic features of speed and rate (<prosody>), and how to change the speaking

SSML 1.0 [21] continued to be standardized to promote the use of SSML to more international languages, in particular Asian and Indian languages. Three workshops were held to encourage local companies and universities to propose features to be

A new standard SSML 1.1 [24] was released in September 2010. See Appendix F of [24] for details on the changes. Among them, a < token> element was introduced for languages where the whitespace has peculiar behavior, such as in Mandarin,

Version 1.0, SSML 1.0 [20], which was released in March 2004.

**54**

**Figure 4.**

#### **Figure 5.** *A simple SSML document.*

#### **3.4 Pronunciation lexicon: PLS 1.0**

Both speech grammars and synthesized prompts can require customizing the pronunciation of words in a specific application domain. This is often done by adding a user lexicon. The Pronunciation Lexicon Specification (PLS 1.0 [25]) was created to support the definition of a standard lexicon fully interoperable with SRGS 1.0 and SSML 1.0/1.1. PLS 1.0 became a W3C Recommendation in October 2008.

A PLS document is a container of entries, <lexeme> elements, with a textual part described by the <grapheme> element and with textual replacements provided by <alias> elements or phonetic transcriptions by <phoneme> elements. There can be multiple pronunciations to accommodate different ways to speak a word/token, or for a different spelling for the same pronunciation.

A simple PLS 1.0 document example for a flight application is shown in **Figure 6**. For "Alitalia" and "Lufthansa," the pronunciations inside the <phoneme> element are given in IPA (International Phonetic Alphabet) [34]—a standard way to express the pronunciations for all spoken human languages. Moreover, the two lexemes have a double pronunciation; the first is the normal English one, while the second is closer to their original language (Italian and German, respectively) as spoken by a native speaker of that language. The prefer attribute indicates which pronunciation has to be selected for TTS rendering. For ASR, all the pronunciations will be taken into account.

#### **3.5 Call control—CCXML 1.0**

Another language defined by the W3C VBWG targets programming the call control of a voice browser in an innovative way. An XML markup language was developed to define handlers for telephony events generated by a telephone connection or a VoIP SIP interaction. The Voice Browser Call Control (CCXML 1.0) [26] language was designed to allow a very efficient implementation completely based upon events and handlers to avoid creating any latency that might impact the underlying signaling.

#### **Figure 6.**

*PLS 1.0 document for flight applications.*

A CCXML engine is also able to send and receive events through an HTTP/ HTTPS connector, which allows for the generation of outbound calls from a Web application and for monitoring calls and conferences via a Web interface.

CCXML 1.0 addresses both simple tasks of call handling (see **Figure 7**), as well as complex ones, such as conditional call handling, conferencing, coaching, etc. Each CCXML document describes transitions to handle specific events. In **Figure 7**, a "connection.alerting" event (incoming call) is accepted by the underlying telephony or VoIP layer, a VoiceXML dialog is started when the "connection.connected" event is received, and then the CCXML processor waits until either the caller disconnects ("connection.disconnect") or the VoiceXML dialog exits ("dialog.exit"). These are simple actions performed during telephony calls, both TDM and VoIP.

While working on the definition of CCXML 1.0, which became a W3C Recommendation in July 2011, the W3C VBWG decided to start another effort to define a state-chart language to generalize the ideas behind CCXML 1.0. This new specification is State Chart XML (SCXML): State Machine Notation for Control Abstraction (SCXML 1.0 [26]), and it can be used as the key component to control a generalized interaction in a multimodal interface. SCXML 1.0 is an XML markup language that provides a generic state-machine-based execution environment inspired by Harel state charts [35].

#### **3.6 IETF protocols: MRCPv1 and v2**

The implementation of voice browsing relies on other standards and protocols, the web architecture, with XML documents, namespaces, caching policies to start with, and obviously the HTTP/HTTPS protocols. All these are at the core of the W3C VBWG standards. However, the Internet Task Force Initiative (IETF) [36] was working on needed protocols.

The Media Resource Control Protocol (MRCP), whose initial draft was proposed by CISCO, SpeechWorks, and Nuance, defines the requests, responses, and events to

**57**

standards, see [39].

**Figure 7.**

**4. W3C MMIWG standards**

*Basic handling of incoming calls with CCXML.*

*Speech Standards: Lessons Learnt*

*DOI: http://dx.doi.org/10.5772/intechopen.93134*

control resources of general speech engines, such as ASR and TTS and even speaker verification to enable a distributed and scalable architecture. The initial draft was standardized by IETF as MRCPv1 (RFC 4463 [37]), and it was largely implemented by the industry. The protocol was based on Real-Time Transport Protocol (RTP) for media transport and RTSP (Real Time Streaming Protocol) for controlling speech resources. In the meantime, standardization continued to MRCPv2, which was instead

based on SIP (Session Initiation Protocol) for signaling and SDP (Session Description Protocol) for negotiating and exchanging capabilities. In November 2012, the standardization was completed (RFC 6787 [38]), and it enabled the control of new resources for recording, speaker verification, and identification. For a complete description of MRCPv2 and its relationship with W3C VBWG

The companion working group, Multi-Modal Interaction Working Group (MMIWG), led by Deborah Dahl was attended by almost the same companies attending VBWG. The goal of MMIWG was to extend the scope of standardization beyond the voice or typed input to embrace a much larger set of modalities, such as touch, gesture, emotions, and haptics both as input and output devices for a system. The major achievements of the W3C MMIWG were the following standards:

• Ink Markup Language, InkML [40], is designed to represent the input of handwriting by a stylus or a finger. In addition to representing traces, InkML offers a rich set of metadata that preserve the appearance of the original input

(i.e., color, width, orientation, timing, etc.).

*Speech Standards: Lessons Learnt DOI: http://dx.doi.org/10.5772/intechopen.93134*

*Human 4.0 - From Biology to Cybernetic*

A CCXML engine is also able to send and receive events through an HTTP/ HTTPS connector, which allows for the generation of outbound calls from a Web

CCXML 1.0 addresses both simple tasks of call handling (see **Figure 7**), as well as complex ones, such as conditional call handling, conferencing, coaching, etc. Each CCXML document describes transitions to handle specific events. In **Figure 7**, a "connection.alerting" event (incoming call) is accepted by the underlying telephony or VoIP layer, a VoiceXML dialog is started when the "connection.connected" event is received, and then the CCXML processor waits until either the caller disconnects ("connection.disconnect") or the VoiceXML dialog exits ("dialog.exit"). These are simple actions performed during telephony calls, both TDM and VoIP. While working on the definition of CCXML 1.0, which became a W3C Recommendation in July 2011, the W3C VBWG decided to start another effort to define a state-chart language to generalize the ideas behind CCXML 1.0. This new specification is State Chart XML (SCXML): State Machine Notation for Control Abstraction (SCXML 1.0 [26]), and it can be used as the key component to control a generalized interaction in a multimodal interface. SCXML 1.0 is an XML markup language that provides a generic state-machine-based execution environment

The implementation of voice browsing relies on other standards and protocols, the web architecture, with XML documents, namespaces, caching policies to start with, and obviously the HTTP/HTTPS protocols. All these are at the core of the W3C VBWG standards. However, the Internet Task Force Initiative (IETF) [36] was

The Media Resource Control Protocol (MRCP), whose initial draft was proposed by CISCO, SpeechWorks, and Nuance, defines the requests, responses, and events to

application and for monitoring calls and conferences via a Web interface.

**56**

**Figure 6.**

*PLS 1.0 document for flight applications.*

inspired by Harel state charts [35].

working on needed protocols.

**3.6 IETF protocols: MRCPv1 and v2**

#### **Figure 7.** *Basic handling of incoming calls with CCXML.*

control resources of general speech engines, such as ASR and TTS and even speaker verification to enable a distributed and scalable architecture. The initial draft was standardized by IETF as MRCPv1 (RFC 4463 [37]), and it was largely implemented by the industry. The protocol was based on Real-Time Transport Protocol (RTP) for media transport and RTSP (Real Time Streaming Protocol) for controlling speech resources.

In the meantime, standardization continued to MRCPv2, which was instead based on SIP (Session Initiation Protocol) for signaling and SDP (Session Description Protocol) for negotiating and exchanging capabilities. In November 2012, the standardization was completed (RFC 6787 [38]), and it enabled the control of new resources for recording, speaker verification, and identification.

For a complete description of MRCPv2 and its relationship with W3C VBWG standards, see [39].


**1. Introduction**

Magnetic resonance imaging (MRI) is the best imaging technique for the evaluation of the neurologic system due to the ability to diagnose central nervous system (CNS) diseases in both humans and animals, especially diseases affecting cerebral white matter (CWM), providing in vivo markers for disease severity or response to therapy and shedding light on progression and recovery processes. The broad spectrum of magnetic resonance contrast mechanisms makes MRI one of the most important and widely used imaging tool for diagnosis in the CNS. However, one of the disadvantages of conventional MRI is that it does not allow the visualization of the cerebral White matter constituent fiber tracts and their connectivity in the brain in vivo [1].

© 2016 The Author(s). Licensee InTech. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. © 2016 The Author(s). Licensee InTech. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Previously, the anatomy of the cerebral white matter tracts could only be studied by postmortem dissection or through invasive methods, and said methods could only reveal a few tracts in vivo (neurosurgery) and no tracts could be observed in vivo via conventional imaging studies [1–6].

Diffusion tensor tractography (DTT) is a useful noninvasive imaging technique that can identify and represent fiber tracts of the cerebral white matter and their connections in the brain in vivo [1]; also, it can give us information that cannot be achieved by conventional anatomical MRI or histology. In addition to displaying specific cerebral white matter fiber tracts, this technique can also improve the quantification of diffusion characteristics within these fibers [1, 7–9].

DTT provides a three-dimensional representation of diffusion tensor imaging (DTI), and data can be displayed on a colored map obtained from information on the directionality of the movement of water molecules along the main fiber tracts of cerebral white matter [1].

#### **1.1. Diffusion tensor imaging**

DTI uses the property of the water diffusion anisotropy in axonal fibers allowing the analysis and tracking of said fibers in the brain [1, 4, 10, 11].

DTI permits the exploration of microstructural tissue features through the observation of water molecular diffusion, thus furnishing information about the anatomy, microstructural features, and damage of the main brain bundles, useful in several pathological animal models. DTIbased tractography permits the virtual reconstruction of the white matter fiber bundles invivo, following the principal diffusion direction [12, 13].

This technique is commonly used in human medicine to study the anatomy and maturation of the normal, aging brain, but it also can be used to help diagnose neurological conditions, including brain ischemia, multiple sclerosis, diffuse axonal injury, epilepsy, metabolic disorders, certain mental illnesses, and brain tumors, as well as establish a prognosis for patients with these conditions [14, 15]. Though DTI has been extensively used to investigate brains of the dogs ex vivo [11]; there is only one report of the in vivo use of DTT to study cerebral white matter fiber tracts in dogs [1].

The ability to trace cerebral white matter fibers in the dog generates a number of opportunities for potential clinical applications, and has both diagnostic and prognostic [1].


*2.3.2 National level*

Cultivating cotton by adopting closer spacing with the available cultivars and with those having short branches was conceived and implemented in India since sixties of the 20th century. Visits, exposures, dialogs and discussions could improvise the learning and the team's visit to Brazil had sown the idea of researching on High Density Planting System (HDPS) and the churned idea was implemented by reorienting research through All India Coordinated Cotton Improvement Project (AICCIP) and through other schemes like Technology Mission of Cotton (TMC) or National Food Security Mission (NFSM) in India [42]. It is reiterated that the success of HDPS at varied locations solely depends on the availability of genotypes, appropriate spacing and nutrition for adopting more plants per hectare to achieve more productivity per unit area and sound pest management criteria. This necessitates the evaluation of available genotypes in varied spacing and nutrition level to the incidence of insect pests. As farmers tend to grow more Bt which are normally of spreading in nature, evaluation of both Bt and not Bt compact genotypes for

their suitability to HDPS is the need of the hour. The adoption of HDPS along with better genotype and fertilizer management is a viable approach to break the current stagnation of yield.

More the synthetic fertilizer application, especially nitrogen (N) fertilizer, more the serious insect herbivores occurrence and crop damage from these insects by reducing plant resistance, the concept which has been conceived clearly [43, 44]. Reducing fertilizer applications can reduce the production costs for cotton growers, as well as nitrogen (N) leaching into the soil and contamination of surface and ground water, but altered N fertilization may also affect pests and their natural enemies [45]. Occurrence of insects and their abundance are heavily dependent on the micro climate available in the system which is primarily based on the biomass production by the plants and their nearness (spacing). Hence, it is utmost important to study the pest dynamics under closer spacings as well as increased levels of nitrogen applications.

A study taken up using GSHV-01/1338 and GBHV-164 genotypes among others revealed their ability as promising genotypes of the region suited to high density planting system due to its compact nature [42]. At two levels of closer spacings (60x15 and 45x15 cm) against the normal spacing of 120x45 cm along with two increased level of nitrogen application (i.e. 125 and 150% of RDN), these two genotypes performed better. The studies were carried out in factorial randomized block design *Kharif* 2013–2014. Closer spacings (45x15 and 60x15 cm) attracted more thrips as compared to the recommended spacing (120x45 cm). The mean population of thrips was found significantly high on GBHV-164 than GSHV-01/1338. Higher dose of nitrogen application on crop (125 and 150% RDN) attracted more thrips as compared to 100 per cent recommended dose of nitrogen (RDN).

[46] by quoting a field experiment that was conducted to study the mean incidence of major cotton insect pests during two consecutive seasons *i.e*. during *kharif*, 2010–2011 and 2011–2012 at CICR, Nagpur under high density planting system (HDPS) using different genotypes of *G. hirsutum* with different spacings indicated that pest incidence was not altered by closer spacing. The main objectives of the work was to identify lines of *G. hirsutum* which have less infestation of major insect pests under HDPS system and to investigate whether the incidence is influenced by plant density. In 2010–2011, the minimum mean population of leafhopper was observed in NISC-50 (1.82 nymphs / 3 leaves /plant) which was grown at a spacing of 45x13.5 cm followed by PKV-0811 (1.91 nymphs /3 leaves/plant) grown at a spacing of 45x13.5 cm and these genotypes are significantly superior over the others. The injury grade was I in both NISC-50 and PKV-081 genotypes. The mean per cent square damage was low in CNH-120 MB (2.76%) followed by PKV-081 (3.82%), both being statistically on par with each other and significantly superior over other genotypes. The mean pink bollworm population was low on PKV-081 (2.53 larvae/25 green bolls). The lowest per cent locule damage due to pink boll worm was noticed on PKV-081 (8.48%). However, the performance of genotypes and geometry against all the insect pests in 2011–2012 was not significantly different leading to a conclusion that pest incidence was not altered by closer spacing.

[47] reported in a study undertaken during 2015–2016 on High density planting demonstrations (50) which were taken up in farmers' fields at varied close spacings (75x10 and 90x10cm) with available compact genotypes (Suraj and G.Cot.16) compared to normal spacing (120x45 cm) under Insecticide Resistance Management (IRM) umbrella in rainfed regions of Bharuch district. Aphids, thrips and leafhopper were found above ETL whereas whitefly and mealybug were found below ETL. The mean larval population of pink bollworm was 4.41 and 3.14 larvae/20 green bolls in Suraj and G.Cot.16 spaced at closed spacings respectively. The pink bollworm population was 2.51 and 2.68 larvae/20 green bolls in Bt-IRM and non IRM

#### *High Density Planting System of Cotton in India: Status and Breeding Strategies DOI: http://dx.doi.org/10.5772/intechopen.94905*

plots respectively. Suraj variety spaced at 75x10 and 90x10 cm required 4.21 and 3.33 sprays and G.Cot.16 spaced at 75x10 and 90x10 cm required 4.40 and 3.60 sprays against sucking pests and 2.37 and 2.38 and 3.20 and 2.40 sprays against bollworms respectively as against 5.00 and 5.60 sprays against sucking pests and 2.00 and 3.80 sprays against bollworms in Bt-IRM and Bt-Non IRM cotton respectively. These results suggest the need for excessive sprays in ultra closer spacing than the normal closer spacing for both the cultivars. The net return was found higher in G.Cot.16 HDPS at both the spacing (Rs. 22,966 and 17,456/acre) than the Suraj HDPS (Rs. 16,461 and 8235/acre). The net return for Bt-IRM farmers was higher (Rs.21527/ acre) than non IRM-Bt farmers (Rs. 17,919/acre). Thus, it has been concluded that HDPS offer viable option to increase productivity especially under rainfed region.

The cotton crop is attacked by 1326 species of insect pests throughout the world, of which about 130 different species of insects and mites found to devour cotton at different stages of crop growth in India. Among the bollworms, pink bollworm assumed major pest status in recent past [48]. The pink bollworm, *Pectinophora gossypiella* (Saunders), a pest which received more attention in almost all the cotton growing states of India (except Tamil Nadu as of now), is identified as the most destructive pest of cotton and causes 2.8 to 61.9 per cent loss in seed cotton yield, 2.1 to 47.1 per cent loss in oil content and 10.7 to 59.2 per cent loss in normal opening of bolls [49]. Locule damage was noted to an extent of 55 per cent and 35–90 per cent reduction in seed cotton yield has been reported by [50, 51] estimated the yield loss to an extent of 6525 MT annually.

Losses caused by pests vary by 10–30% depending on the intrinsic genetic factors and its rigidity in expressing the inherent resistance. Pests are supposed to evolve in a short and strategic cycle to circumvent the problems being arisen and judicious use of insecticides along with physical, biological control methods is the need of the hour. Ignoring pests can lead to complete crop failure. In the overall crop protection program under the National Agricultural Policy, The Government's IPM is a time-tested, eco-friendly approach, socially acceptable and economically viable that is widely accepted across the country. Appropriate control measures should be taken when insect populations cross the ETL [52].

#### **2.4 Diseases management**

High density planting which entails closer planting may have every chance of microclimate getting altered due to which the propensity of infectious diseases in cotton may also vary. The high density planting in cotton is a recent phenomena which opened avenues for research in various domains including plant pathology. The plant pathologists have been trying to understand the nexus between the incidence of various cotton diseases and the change in microclimate of the plant coupled with external atmosphere [53, 54].

The life cycle of pathogens is amenable for changes in line with the changes in plant canopy and the microclimate mediated through weather parameters. Space between plants and rows is bound to have a say in the mode of dispersal, the intensity of infection and the production of secondary inoculum of plant pathogens. Cotton, being a commercial crop, is no exception to this phenomena of infection and the high density planting in cotton need to be carefully contemplated taking into account the changed plant geometry and the corresponding incidence of cotton diseases. Despite the research on influence of high density planting in cotton on the incidence of diseases is in the nascent stage, an attempt has been made to take stock of striking developments in the management of important cotton diseases in the succeeding pages.

The major diseases of cotton which are prevalent in most part of the cotton growing countries in the world were reported to inflict a damage ranging from 10 to 30% and it may be more when favorable conditions prevail for the spread of the pathogens which culminates in cotton farmers spending huge cost to keep the biotic stress under control [55].

Fungal diseases are predominant in cotton followed by very few bacterial and viral diseases. The prominent bacterial disease which inflict major damage in cotton crop is bacterial blight, caused by *Xanthomonas citri* pv. *malvacearum* [56]. Abundant literature is available on major fungal diseases of cotton namely *Fusarium* wilt caused by *Fusarium oxysporum* f. sp. *vasinfectum*, *Verticillium* wilt caused by *Verticillium dahliae*, anthracnose caused by *Colletotrichum gossypii*, Ramularia gray mildew caused by *Mycosphaerella areola*, root rots caused by *Rhizoctonia solani* and *R. bataticola*, leaf blight caused by *Alternaria macrospora* and leaf spot caused by *Cercospora gossypina* [57–62]. The cotton leaf curl disease is the only virus disease documented in cotton which belongs to the genus Begomovirus and transmitted by insect vectors [63].

Plethora of studies were conducted for the management of cotton diseases which reported solitary or combination of management practices to control them. Besides chemical control, significant research work has been carried out on the biological control of cotton diseases [64–67]. Similarly, there were prominent studies on use of plant extracts [68–70] and essential oil [71, 72] for the management of cotton diseases. Cultural methods [73] and organic amendments [74] also form part of the strategy to control cotton diseases.

In the recent past, several research studies have documented the efficacy of Endophytic bacteria [75] in suppressing the incidence of cotton diseases. Molecular level studies namely transcriptomic, proteomic and metabolomic studies [76–78] and studies on gene editing [79] were on the rise for the past two decades. As there are several methods and molecules have been designed for effecting the control of diseases, their role in containing the diseases that occur under HDPS is in infant stage as the genotypes which have been evaluated at Coimbatore were not found to have adequate disease expressions under HDPS.

#### *2.4.1 Soil borne diseases*

Soil borne fungal diseases of cotton namely damping off, root rot and wilt have been reported to cause extensive damage in cotton crop. Juxtaposing chemical control with biological method, the latter was found to be effective which is evidenced from the finding of [80–82] who reported that the combined application of *T. harzianum* and *P. lilacinus* showed the best result by inhibiting the growth of pathogen than alone. A recent study of [83] reported that Trichodel®, based on *Trichoderma* spp., reduced the incidence and severity of wilt caused by *F. oxysporum* f. sp. *vasinfectum*. Besides, a score of agronomic practices namely fine tilth of the soil, adjusting sowing season, crop rotation, soil solarization, amending soil for altering pH of the soil and use of resistant varieties have been reported to reduce the incidence of soil borne diseases. Biological control of *V. dahliae* in cotton with a mixture of lignin and *Trichoderma viride* [84] has been reported. Thus, biological control of soil borne diseases is the viable option which had been arrived by various authors in the normal cotton growing situations. However, the same might hold good under HDPS also.

#### *2.4.2 Foliar diseases*

Among the foliar diseases, Alternaria leaf spot, gray mildew, boll rot, rust, anthracnose and bacterial leaf blight were reported in cotton and they were reported to inflict damage significantly. The chemical fungicides mancozeb (0.3%), propiconazole (0.1%), propineb (0.3%) were found more effective against

#### *High Density Planting System of Cotton in India: Status and Breeding Strategies DOI: http://dx.doi.org/10.5772/intechopen.94905*

*Alternaria macrospora*, propiconazole (0.1%) and copper oxychloride (0.25%) against *Myrothecium roridum* [85, 86]. Moreover, a decadal analysis of *Alternaria* occurrence among the various genotypes at Coimbatore indicated that Bunny Bt cotton, NCEHBT, Dhannu BGII and 1037 BGII genotypes were found to be more susceptible and the disease incidence ranged from 0.5 to 10.53 PDI compared to the types which are resistant/field tolerant. In addition, the sowing taken up during 29th–30th Standard Meteorological Week (SMW) resulted in lesser incidence of the disease irrespective of the cultures evaluated [87].

Among the biocontrol agents studied, *Pseudomonas fluorescens* strains and *Bacillus subtilis* and the botanicals derived from *Azadirachata indica*, *Lantana camera*, *Calotropis procera*, *Ocimum sanctum*, *Allium cepa* and *Allium sativum* have been reported to significantly reduce the mycelial growth of the pathogenic fungus [88]. The methanol extracts of *Polyalthia longifolia* and *Terminalia chebula* and chloroform extract of *Zingiber officinale*, *Datura alba*, *Moringa olifera*, *Azadirachta indica* and *Syzgyium cumini* have showed significant biological control of cotton bacterial blight in greenhouses and in fields [89, 90]. This indicates that principles available in various plants are having sizable influence in containing the growth of disease causing micro organisms.

## *2.4.3 Viral diseases*

Among the viral diseases infecting cotton, cotton leaf curl virus and tobacco streak virus are important. The TSV disease was reported to be spread through mechanical means, infected seeds and through thrips species. Parthenium, a widely distributed and symptom less carrier of TSV, plays a major role in perpetuation and spread of the disease [91–95].

A study carried out by [96] uncovered the application of *Bacillus* species which possess diverse anti microbial peptide (AMP) genes which are responsible for the biosynthesis of antibiotics like iturin, bacilysin, bacillomycin, fengycin, surfactin, mersacidin, ericin, subtilin, subtilosin, and mycosubtilin in curtailing the infection of TSV. Genetic Engineering studies to evolve transgenic cotton using an antisense RNA approach [97] could be a potential option for managing the disease. Interestingly, transgenic cotton plants that over express miR166 also show potential in reducing *Bemisia tabaci* populations and, more importantly, the spread of whitely transmitted plant viruses [98, 99]. Gene editing technology *i.e.,* CRISPR/Cas9 system has recently been used to confer molecular immunity against several eukaryotic viruses, including cotton DNA geminiviruses [100].


**Acknowledgements**

This study was partially supported by the First Bank of Toyama Scholarship foundation and a Grants-in-Aid for Scientific Research (No. 21671003). We are most grateful to Ana Pantar and Book Editors for their editorial advices and cooperation.


**Bone Marrow Derived Pluripotent Stem Cells in Ischemic Heart Disease: Bridging the Gap Between Basic Research and Clinical Applications**

Ahmed Abdel-Latif1, Ewa Zuba-Surma2 and Mariusz Z. Ratajczak3 *1Gill Heart Institute and Division of Cardiovascular Medicine, University of Kentucky, and Lexington VA Medical Center, Lexington, KY 2Department of Medical Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Cracow, 3Stem Cell Biology Institute, James Graham Brown Cancer Center, University of Louisville, Louisville, KY,*

*1,3USA 2Poland*

### **1. Introduction**

424 Advances in Hematopoietic Stem Cell Research

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real-time TaqMan PCR for routine quantitation of cytomegalovirus DNA in crude leukocyte lysates from stem cell transplant patients. *J Virol Methods* Vol.110, No.1, The prevalence of ischemic heart disease and acute myocardial infarction (AMI) has increased to alarming rates in the United States and the western world (Roger *et al.,* 2011). Patients who survive the initial AMI suffer ischemic cardiomyopathy (ICM) which is often complicated by high mortality and poor overall prognosis (Braunwald *et al.,* 2000; McMurray *et al.,* 2005). Despite significant advances in medical therapy and revascularization strategies, the prognosis of patients with AMI and ischemic cardiomyopathy remains dismal (Levy D *et al.,* 2002; Roger VL *et al.,* 2004). The last decade has demonstrated significant progress and rapid translation of myocardial regenerative therapies particularly those utilizing stem cells isolated from adult tissues (Abdel-Latif *et al.,* 2007).

Studies examining the potential therapeutic use of bone marrow (BM)-derived cells in myocardial regeneration have overshadowed the growing evidence of innate cardiac reparatory mechanisms. Several studies have demonstrated the capability of cardiomyocytes to replenish through poorly understood innate mechanisms. Follow up of cardiac transplantation patients have demonstrated continuous replenishment of cardiomyocytes by recipient's derived cells through poorly understood mechanisms (Quaini *et al.,* 2002). There is growing evidence that BM-derived cells are responsible, at least in part, for organ chimerism including cardiomyocyte chimerism (de Weger *et al.,* 2008; Deb *et al.,* 2003). Animal studies have confirmed this to be a dynamic process responding to significant injury such as myocardial infarction and peaks in the peri-infarct zone (Hsieh *et al.,* 2007). Although this process appears to be robust enough to achieve the renewal of approximately 50% of all cardiomyocytes in the normal life span, very little is known about its underpinnings (Bergmann *et al.,* 2009).

Bone Marrow Derived Pluripotent Stem Cells in

pluripotent potential.

Ischemic Heart Disease: Bridging the Gap Between Basic Research and Clinical Applications 427

have been also isolated from skeletal muscle and other tissues (Abdel-Latif *et al.,* 2008a). However, it is conceivable that different investigators have isolated, using different methods, the same or very similar populations and named them differently. It is also possible that these populations at least in part contain VSELs which might explain their

**Panel A:** Gating strategy for isolating human cord blood (CB)-derived VSELs. Morphology of total CBderived nucleated cells is shown on dot-plot representing FSC and SSC parameters related to cell size and granularity/ complexity, respectively. All objects larger than 2m are enclosed in region R1 and further visualized on histogram showing the expression of markers of mature hematopoietic cells (lineage markers; Lin). Cells not expressing differentiated hematopoietic markers (Lin- in region R2) are then analyzed for CD34 and CD45 expression. VSELs are identified as CD45-/Lin-/CD34+ cells (region

**Panel B**: Sorting of murine bone marrow (BM)-derived VSELs. Morphology of total murine BM-derived nucleated cells is shown on dot-plot presenting FSC and SSC parameters and all objects in range of 2- 10m in diameter are included in region R1. Lymphocytic cells including stem cell fraction is further analyzed for Sca-1 and differentiated hematopoietic lineages markers (Lin) expression and only Sca-1+/Lin- cells are included in region R2. Cells from this region are further seperated based on CD45 expression. Murine VSELs are distinguished as CD45-/Lin-/Sca-1+ cells (region R3), while HSCs as

Fig. 1. Strategy for flow cytometric analysis of human and murine Very Small Embryonic-Like and hematopoietic stem cells. Briefly, BM is flushed from the femurs and tibias. Nucleated cells are isolated by lysis of red blood cells and cells are then gated on based on the cell size (>2 m). Of note, lysis is preferred for isolating VSELs rather than Ficoll gradient that we have shown to lose some of the VSELs due to their small size.

R3), while hematopoietic stem cells (HSCs) as CD45+/Lin-/CD34+ cells (region R4).

CD45+/Lin-/Sca-1+ cells (region R4).

Complex innate reparatory mechanisms are initiated by myocardial ischemia interacting with different elements of the immune system, the infarcted myocardium and bone marrow stem cells, culminating in BM-stem and progenitor cells' (SPCs) mobilization as we and others have demonstrated (Abdel-Latif *et al.,* 2010; Walter *et al.,* 2007; Wojakowski *et al.,* 2009). However, very little is known about the mechanisms and clinical significance of this mobilization. Animal studies show that mobilized BM-derived cells (BMCs) repopulate the infarct border, however the significance of this mobilization is unclear given the low rate of their differentiation into cardiomyocytes (Fukuhara *et al.,* 2005).