Effects of Transparent Testa8 (TT8) gene and Homeobox12 (HB12) gene silencing in alfalfa (Medicago sativa L.) on molecular structure spectral profile in relation to energy, degradation, and fermentation characteristics in ruminant systems

Alfalfa (Medicago sativa L.) is a legume forage that is widely cultivated owing to its high biomass yield and favorable nutrient values. However, alfalfa contains relatively high lignin, which limits its utilization. Downregulation of two transcriptional factors, Transparent Testa8 (TT8) and Homeobo...

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Main Authors: Yaogeng Lei, Abdelali Hannoufa, Peiqiang Yu
Format: Article
Language:English
Published: KeAi Communications Co., Ltd. 2023-09-01
Series:Animal Nutrition
Subjects:
Online Access:http://www.sciencedirect.com/science/article/pii/S240565452300029X
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author Yaogeng Lei
Abdelali Hannoufa
Peiqiang Yu
author_facet Yaogeng Lei
Abdelali Hannoufa
Peiqiang Yu
author_sort Yaogeng Lei
collection DOAJ
description Alfalfa (Medicago sativa L.) is a legume forage that is widely cultivated owing to its high biomass yield and favorable nutrient values. However, alfalfa contains relatively high lignin, which limits its utilization. Downregulation of two transcriptional factors, Transparent Testa8 (TT8) and Homeobox12 (HB12), has been proposed to reduce lignin content in alfalfa. Therefore, silencing of TT8 (TT8i) and HB12 (HB12i) in alfalfa was achieved by RNAi technology. The objective of this project was to determine effect of gene modification through silencing of TT8 and HB12 genes in alfalfa plants on lignin and phenolic content, bioenergic value, nutrient supply from rumen degradable and undegradable fractions, and in vitro ammonia production in response to the silencing of TT8 and HB12 genes in alfalfa. All gene silenced alfalfa plants (5 TT8i and 11 HB12i) were grown under greenhouse conditions with wild type as a control. Samples were analyzed for bioactive compounds, degradation fractions, truly digestible nutrients, energetic values and in vitro ammonia productions in ruminant systems. Furthermore, relationships between physiochemical, metabolic and fermentation characteristics and molecular spectral parameters were determined using vibrational molecular spectroscopy. Results showed that the HB12i had higher lignin, while TT8i had higher phenolics. Both silenced genotypes had higher rumen slowly degraded carbohydrate fractions and truly digestible neutral detergent fiber, but lower rumen degradable protein fractions. Moreover, the HB12i had lower truly digestible crude protein, energetic values and ammonia production compared with other silenced genotypes. In addition, in relation to the nutritive values of alfalfa, structural carbohydrate parameters were negatively correlated, whereas alpha/beta ratio in protein structure was positively correlated. Furthermore, good predictions were obtained for degradation of protein and carbohydrate fractions and energy values from molecular spectral parameters. In conclusion, silencing of the TT8 and HB12 genes decreased protein availability and increased fiber availability. Silencing of the HB12 gene also increased lignin and decreased energy and rumen ammonia production. Moreover, nutritional alterations were closely correlated with molecular spectral parameters. Therefore, gene modification through silencing the TT8 and HB12 genes in alfalfa influenced physiochemical, metabolic and fermentation characteristics.
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spelling doaj.art-6648eab07e0e4c52b7339de3840069af2023-09-30T04:55:08ZengKeAi Communications Co., Ltd.Animal Nutrition2405-65452023-09-01147987Effects of Transparent Testa8 (TT8) gene and Homeobox12 (HB12) gene silencing in alfalfa (Medicago sativa L.) on molecular structure spectral profile in relation to energy, degradation, and fermentation characteristics in ruminant systemsYaogeng Lei0Abdelali Hannoufa1Peiqiang Yu2Department of Animal and Poultry Science, College of Agriculture and Bioresources, University of Saskatchewan, 51 Campus Drive, Saskatoon, SK S7N5A8, CanadaLondon Research and Development Centre, Agriculture and Agri-Food Canada, 1391 Sandford Street, London, ON N5V 4T3, CanadaDepartment of Animal and Poultry Science, College of Agriculture and Bioresources, University of Saskatchewan, 51 Campus Drive, Saskatoon, SK S7N5A8, Canada; Corresponding author.Alfalfa (Medicago sativa L.) is a legume forage that is widely cultivated owing to its high biomass yield and favorable nutrient values. However, alfalfa contains relatively high lignin, which limits its utilization. Downregulation of two transcriptional factors, Transparent Testa8 (TT8) and Homeobox12 (HB12), has been proposed to reduce lignin content in alfalfa. Therefore, silencing of TT8 (TT8i) and HB12 (HB12i) in alfalfa was achieved by RNAi technology. The objective of this project was to determine effect of gene modification through silencing of TT8 and HB12 genes in alfalfa plants on lignin and phenolic content, bioenergic value, nutrient supply from rumen degradable and undegradable fractions, and in vitro ammonia production in response to the silencing of TT8 and HB12 genes in alfalfa. All gene silenced alfalfa plants (5 TT8i and 11 HB12i) were grown under greenhouse conditions with wild type as a control. Samples were analyzed for bioactive compounds, degradation fractions, truly digestible nutrients, energetic values and in vitro ammonia productions in ruminant systems. Furthermore, relationships between physiochemical, metabolic and fermentation characteristics and molecular spectral parameters were determined using vibrational molecular spectroscopy. Results showed that the HB12i had higher lignin, while TT8i had higher phenolics. Both silenced genotypes had higher rumen slowly degraded carbohydrate fractions and truly digestible neutral detergent fiber, but lower rumen degradable protein fractions. Moreover, the HB12i had lower truly digestible crude protein, energetic values and ammonia production compared with other silenced genotypes. In addition, in relation to the nutritive values of alfalfa, structural carbohydrate parameters were negatively correlated, whereas alpha/beta ratio in protein structure was positively correlated. Furthermore, good predictions were obtained for degradation of protein and carbohydrate fractions and energy values from molecular spectral parameters. In conclusion, silencing of the TT8 and HB12 genes decreased protein availability and increased fiber availability. Silencing of the HB12 gene also increased lignin and decreased energy and rumen ammonia production. Moreover, nutritional alterations were closely correlated with molecular spectral parameters. Therefore, gene modification through silencing the TT8 and HB12 genes in alfalfa influenced physiochemical, metabolic and fermentation characteristics.http://www.sciencedirect.com/science/article/pii/S240565452300029XGene silencingHomeobox12 (HB12) geneTransparent Testa8 (TT8) geneAlfalfa (Medicago sativa L.)Ruminant systemFermentation and degradation
spellingShingle Yaogeng Lei
Abdelali Hannoufa
Peiqiang Yu
Effects of Transparent Testa8 (TT8) gene and Homeobox12 (HB12) gene silencing in alfalfa (Medicago sativa L.) on molecular structure spectral profile in relation to energy, degradation, and fermentation characteristics in ruminant systems
Animal Nutrition
Gene silencing
Homeobox12 (HB12) gene
Transparent Testa8 (TT8) gene
Alfalfa (Medicago sativa L.)
Ruminant system
Fermentation and degradation
title Effects of Transparent Testa8 (TT8) gene and Homeobox12 (HB12) gene silencing in alfalfa (Medicago sativa L.) on molecular structure spectral profile in relation to energy, degradation, and fermentation characteristics in ruminant systems
title_full Effects of Transparent Testa8 (TT8) gene and Homeobox12 (HB12) gene silencing in alfalfa (Medicago sativa L.) on molecular structure spectral profile in relation to energy, degradation, and fermentation characteristics in ruminant systems
title_fullStr Effects of Transparent Testa8 (TT8) gene and Homeobox12 (HB12) gene silencing in alfalfa (Medicago sativa L.) on molecular structure spectral profile in relation to energy, degradation, and fermentation characteristics in ruminant systems
title_full_unstemmed Effects of Transparent Testa8 (TT8) gene and Homeobox12 (HB12) gene silencing in alfalfa (Medicago sativa L.) on molecular structure spectral profile in relation to energy, degradation, and fermentation characteristics in ruminant systems
title_short Effects of Transparent Testa8 (TT8) gene and Homeobox12 (HB12) gene silencing in alfalfa (Medicago sativa L.) on molecular structure spectral profile in relation to energy, degradation, and fermentation characteristics in ruminant systems
title_sort effects of transparent testa8 tt8 gene and homeobox12 hb12 gene silencing in alfalfa medicago sativa l on molecular structure spectral profile in relation to energy degradation and fermentation characteristics in ruminant systems
topic Gene silencing
Homeobox12 (HB12) gene
Transparent Testa8 (TT8) gene
Alfalfa (Medicago sativa L.)
Ruminant system
Fermentation and degradation
url http://www.sciencedirect.com/science/article/pii/S240565452300029X
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